A super-resolution holographic microscope includes a light source configured to emit input light, a diffraction grating configured to split the input light into first diffracted light and second diffracted light, a mirror configured to reflect the first diffracted light, a wafer stage arranged on an optical path of the second diffracted light and on which a wafer is configured to be arranged, and a camera configured to receive the first diffracted light that is reflected by the mirror and the second diffracted light that is reflected by the wafer to generate a plurality of hologram images of the wafer.

A phase distribution is calculated such that modulated light has a predetermined intensity distribution on a target plane and displayed on a phase modulation plane, readout light enters the phase modulation plane so as to generate the modulated light. When calculating the phase distribution, a region on the phase modulation plane is divided into N regions A.sub.1 . . . A.sub.N, with sizes set such that integration values of an intensity distribution in the regions are equal to each other. Further, a region on the target plane is divided into N regions B.sub.1 . . . B.sub.N, with sizes set such that integration values of an intensity distribution in the regions are equal to each other. The phase distribution is calculated by obtaining an optical path length from the region A.sub.n to the region B.sub.n, and determining the phase of the region A.sub.n based on the optical path length.

A method for lens-free imaging of a sample or objects within the sample uses multi-height iterative phase retrieval and rotational field transformations to perform wide FOV imaging of pathology samples with clinically comparable image quality to a benchtop lens-based microscope. The solution of the transport-of-intensity (TIE) equation is used as an initial guess in the phase recovery process to speed the image recovery process. The holographically reconstructed image can be digitally focused at any depth within the object FOV (after image capture) without the need for any focus adjustment, and is also digitally corrected for artifacts arising from uncontrolled tilting and height variations between the sample and sensor planes. In an alternative embodiment, a synthetic aperture approach is used with multi-angle iterative phase retrieval to perform wide FOV imaging of pathology samples and increase the effective numerical aperture of the image.

In situ investigation of microbial life in extreme environments can be carried out with microscopes capable of imaging 3-dimensional volumes and tracking particle motion. A lensless digital holographic microscope is disclosed that provides roughly 1.5 micron resolution in a compact, robust package suitable for remote deployment. High resolution is achieved by generating high numerical-aperture input beams with radial gradient-index rod lenses. The ability to detect and track prokaryotes was explored using bacterial strains of two different sizes. In the larger strain, a variety of motions were seen, while the smaller strain was used to demonstrate a detection capability down to micron scales.

An in-line holographic microscope can be used to analyze on a frame-by-frame basis a video stream to track individual colloidal particles' three-dimensional motions. The system and method can provide real time nanometer resolution, and simultaneously measure particle sizes and refractive indexes. Through a combination of applying a combination of Lorenz-Mie analysis with selected hardware and software methods, this analysis can be carried out in near real time. An efficient particle identification methodology automates initial position estimation with sufficient accuracy to enable unattended holographic tracking and characterization.

In-line holography to create images of a specimen, such as one or more particles dispersed in a transparent medium. Analyzing these images with results from light scattering theory yields the particles' sizes with nanometer resolution, their refractive indexes to within one part in a thousand, and their three dimensional positions with nanometer resolution. This procedure can rapidly and directly characterize mechanical, optical and chemical properties of the specimen and its medium.

Object interference in biological samples generated by lateral shearing interference microscopes is addressed by a shearing microscope slide comprising a periodic structure having alternating reference and sample regions. In some embodiments, the reference regions are configured to provide references that remove sample overlap in a sheared microscopic measurement. A system for generating sheared microscopic measurements is also provided that comprises an inlet configured to receive a sample material, an outlet configured to release a portion of the sample material, and a periodic structure having a plurality of interleaved reference and sample channels. In some cases, the sample channels are configured to accommodate a flow of sample material from the inlet to the outlet and the reference channels are configured to provide references that remove sample overlap in a sheared microscopic measurement.

A method of analyzing a sample receiving a particle of interest, including: defining a reference point located on a first interface of the sample, or at a known distance from the sample, along the optical axis of the optical system; acquiring a reference image transmission of the sample, the object plane of the optical system being located at a known distance from the reference point along an axis parallel to the optical axis of the optical system, and the particle of interest being located outside of the object plane; using the reference image, digitally constructing a series of reconstructed images, each associated with a predetermined offset of the object plane along the optical axis of the optical system; and using the series of reconstructed images, determining the distance along an axis parallel to the optical axis of the optical system, between the particle of interest and the reference point.

A method of generating a color image of a sample includes obtaining a plurality of low resolution holographic images of the sample using a color image sensor, the sample illuminated simultaneously by light from three or more distinct colors, wherein the illuminated sample casts sample holograms on the image sensor and wherein the plurality of low resolution holographic images are obtained by relative x, y, and z directional shifts between sample holograms and the image sensor. Pixel super-resolved holograms of the sample are generated at each of the three or more distinct colors. De-multiplexed holograms are generated from the pixel super-resolved holograms. Phase information is retrieved from the de-multiplexed holograms using a phase retrieval algorithm to obtain complex holograms. The complex hologram for the three or more distinct colors is digitally combined and back-propagated to a sample plane to generate the color image.

Techniques to improve image quality in holography utilizing lenses made from materials with non-quantized anisotropic electromagnetic properties, such as birefringent materials, to advantageously split an incoming beam of light into two coincident beams with different focal lengths that interfere with one another and thus create holograms free of electro-optical or pixelated devices are disclosed for microscopy and other applications. The use of thin birefringent lenses and single crystal alpha-BBO lenses are introduced. Corresponding systems, methods and apparatuses are described.